Journal: In Vitro Cellular & Developmental Biology. Animal
Article Title: Immortalized mouse dental papilla mesenchymal cells preserve odontoblastic phenotype and respond to bone morphogenetic protein 2
Figure Lengend Snippet: Expression of tooth-related genes in the primary and immortalized dental papilla mesenchymal cells. ( A ) Total RNAs from the primary and immortalized cells were extracted and reversely transcribed. The cDNAs were amplified by PCR using specific primers shown in Table 1 . The PCR products were run on 1% agarose gels and stained with ethidium bromide. M DNA marker (Low DNA mass ladder, Invitrogen), Con negative control. ( B ) Immunohistochemistry staining with antibodies against Col1a1, Dlx3, Dmp1, Dsp, Oc, Opn, Osx, and Runx2 in the primary and immortalized murine dental papilla mesenchymal cells. Scale bar = 10 μM. ( C ). Expression of Dsp and Dmp1 proteins in the primary and immortalized mouse dental papilla mesenchymal cells was analyzed by Western blot assay using anti-Dsp and anti-Dmp1 antibodies. Two major fragments of Dmp1 were observed as 57- and 37-kDa, respectively, in a SDS-PAGE gele using anti-Dmp1 antibody. Multiple fragments of Dsp polypeptides were detected in MD10-F2 cells using anti-Dsp antibodiy. Major bands are approximately 120, 65, 60, and 40 kDa as indicated by arrows . β-actin was used as an internal control. Pri and Imm represent the primary and immoratalized dental papilla mesenchymal cells.
Article Snippet: Two-hundred nanograms of DNA (for pSV3 neo plasmid DNA 10 ng) were diluted in a 25-μl polymerase chain reaction (PCR) mix (Sigma–Aldrich).
Techniques: Expressing, Amplification, Polymerase Chain Reaction, Staining, Marker, Negative Control, Immunohistochemistry, Western Blot, SDS Page